TY - JOUR
T1 - A defucosylated anti-CD44 monoclonal antibody 5-mG2a-f exerts antitumor effects in mouse xenograft models of oral squamous cell carcinoma
AU - TAKEI, JUNKO
AU - KANEKO, MIKA K.
AU - OHISHI, TOMOKAZU
AU - HOSONO, HIDEKI
AU - NAKAMURA, TAKURO
AU - YANAKA, MIYUKI
AU - SANO, MASATO
AU - ASANO, TEIZO
AU - SAYAMA, YUSUKE
AU - KAWADA, MANABU
AU - HARADA, HIROYUKI
AU - KATO, YUKINARI
N1 - Funding Information:
This research was supported in part by Japan Agency for Medical Research and Development (AMED) under grant nos. JP20am0401013 (YK), JP20am0101078 (YK), and JP20ae0101028 (YK), and by the Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Scientific Research (KAKENHI) grant nos. 17K07299 (MKK), 19K07705 (YK), and 20K16322 (MS).
Publisher Copyright:
© 2020 Spandidos Publications. All rights reserved.
PY - 2020/11
Y1 - 2020/11
N2 - CD44 is widely expressed on the surface of most tissues and all hematopoietic cells, and regulates many genes associated with cell adhesion, migration, proliferation, differentiation, and survival. CD44 has also been studied as a therapeutic target in several cancers. Previously, an anti-CD44 monoclonal antibody (mAb), C44Mab-5 (IgG1, kappa) was established by immunizing mice with CD44-overexpressing Chinese hamster ovary (CHO)-K1 cells. C44Mab-5 recognized all CD44 isoforms, and showed high sensitivity for flow cytometry and immunohistochemical analysis in oral cancers. However, as the IgG1 subclass of C44Mab-5 lacks antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), the antitumor activity of C44Mab-5 could not be determined. In the present study, we converted the mouse IgG1 subclass antibody C44Mab-5 into an IgG2a subclass antibody, 5-mG2a, and further produced a defucosylated version, 5-mG2a-f, using FUT8-deficient ExpiCHO-S (BINDS-09) cells. Defucosylation of 5-mG2a-f was confirmed using fucose-binding lectins, such as AAL and PhoSL. The dissociation constants (KD) for 5-mG2a-f against SAS and HSC-2 oral cancer cells were determined through flow cytometry to be 2.8x10-10 M and 2.6x10-9 M, respectively, indicating that 5-mG2a-f possesses extremely high binding affinity. Furthermore, immunohistochemical staining using 5-mG2a-f specifically stained the membranes of oral cancer cells. In vitro analysis demonstrated that 5-mG2a-f showed moderate ADCC and CDC activities against SAS and HSC-2 oral cancer cells. In vivo analysis revealed that 5-mG2a-f significantly reduced tumor development in SAS and HSC-2 xenografts in comparison to control mouse IgG, even after injection seven days post-tumor inoculation. Collectively, these results suggest that treatment with 5-mG2a-f may represent a useful therapy for patients with CD44-expressing oral cancers.
AB - CD44 is widely expressed on the surface of most tissues and all hematopoietic cells, and regulates many genes associated with cell adhesion, migration, proliferation, differentiation, and survival. CD44 has also been studied as a therapeutic target in several cancers. Previously, an anti-CD44 monoclonal antibody (mAb), C44Mab-5 (IgG1, kappa) was established by immunizing mice with CD44-overexpressing Chinese hamster ovary (CHO)-K1 cells. C44Mab-5 recognized all CD44 isoforms, and showed high sensitivity for flow cytometry and immunohistochemical analysis in oral cancers. However, as the IgG1 subclass of C44Mab-5 lacks antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), the antitumor activity of C44Mab-5 could not be determined. In the present study, we converted the mouse IgG1 subclass antibody C44Mab-5 into an IgG2a subclass antibody, 5-mG2a, and further produced a defucosylated version, 5-mG2a-f, using FUT8-deficient ExpiCHO-S (BINDS-09) cells. Defucosylation of 5-mG2a-f was confirmed using fucose-binding lectins, such as AAL and PhoSL. The dissociation constants (KD) for 5-mG2a-f against SAS and HSC-2 oral cancer cells were determined through flow cytometry to be 2.8x10-10 M and 2.6x10-9 M, respectively, indicating that 5-mG2a-f possesses extremely high binding affinity. Furthermore, immunohistochemical staining using 5-mG2a-f specifically stained the membranes of oral cancer cells. In vitro analysis demonstrated that 5-mG2a-f showed moderate ADCC and CDC activities against SAS and HSC-2 oral cancer cells. In vivo analysis revealed that 5-mG2a-f significantly reduced tumor development in SAS and HSC-2 xenografts in comparison to control mouse IgG, even after injection seven days post-tumor inoculation. Collectively, these results suggest that treatment with 5-mG2a-f may represent a useful therapy for patients with CD44-expressing oral cancers.
KW - Antibody-dependent cellular cytotoxicity
KW - Antitumor activity
KW - CD44
KW - Complement-dependent cytotoxicity
KW - Monoclonal antibody
KW - Oral cancer
UR - http://www.scopus.com/inward/record.url?scp=85092093801&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85092093801&partnerID=8YFLogxK
U2 - 10.3892/or.2020.7735
DO - 10.3892/or.2020.7735
M3 - Article
C2 - 33000243
AN - SCOPUS:85092093801
SN - 1021-335X
VL - 44
SP - 1949
EP - 1960
JO - Oncology Reports
JF - Oncology Reports
IS - 5
ER -
